Herpetology Notes, volume 14: 777-784 (2021) (published online on 16 May 2021)

Incidence of emerging pathogens in the legal and illegal trade in Spain

Barbora Thumsová1,2, Jaime Bosch3,2,*, and Albert Martinez-Silvestre4

Abstract. are threatened globally and emerging diseases are some of the most important drivers of their catastrophic situation. There is increasing evidence that the international trade in live amphibians is one of the most important mechanisms driving pathogen pollution. Here, we report the presence of Batrachochytrium dendrobatidis (Bd) and Ranavirus in 11% of tested individuals in legal amphibian trade fairs in Spain. Although none of the Bd infected in trade fairs presented disease symptoms, symptoms of ranavirosis were observed in some specimens, which were nonetheless still offered for sale. None of the traders who were selling infected animals showed interest in engaging in collaboration to control infections when offered for free. In addition, a large private urodele collection confiscated by the police in Barcelona comprised a number of illegally wild-caught species. Many confiscated individuals presented signs of poor welfare and several were positive for Bd. Our results indicate the urgent need for implementation of real sanitary regulations or effective legislation governing the practice of trade in living amphibians to prevent pathogen spread in Europe.

Keywords. Batrachochytridium dendrobatidis, chytridiomycosis, Ranavirus, ranavirosis, pet keeping

Introduction al., 2005; Collins and Crump, 2009; Rowley et al., 2010; Pasmans et al., 2017). In fact, of the 5,579 vertebrate Wildlife continues to be threatened by a number of species affected by the global wildlife trade, 9.4% are anthropogenic activities that have compromised the amphibians (Scheffers et al., 2019). Although some integrity and persistence of natural ecosystems over traded amphibian species are monitored by international time (Ceballos et al., 2015; Wilson et al., 2019). In conventions such as the Convention on International recent decades, amphibians have become an iconic Trade in Endangered Species of Wild Fauna and Flora example of species extinction and population extirpation (CITES), regulations for 98% of all amphibian species due to increased human pressure (Skerratt et al., 2007; are not still established (Auliya et al., 2016a). As a Blaustein et al., 2011). As a result, forty-one percent of result, the great majority of traded amphibians are wild- known amphibian species are now listed as threatened, caught, which are mostly harvested in an unsustainable and many are experiencing global population declines manner despite posing risks for population persistence (IUCN, 2020). (Schlaepfer et al., 2005; Auliya et al., 2016b). Among Commercial trade in amphibians for food, pets or other things, the increasing demand from keepers traditional medicine is one of the most prominent to include rare or newly described species in their drivers of species extinction risk globally (Schlaepfer et collections stimulates illegal trade in amphibians (Courchamp et al., 2006; Natusch and Lyons, 2012; Cooney et al., 2015; Auliya et al., 2016b). In fact, the 1 Asociación Herpetológica Española, José Gutiérrez Abascal 2, black market in trade is one of the most lucrative 28006 Madrid, Spain. illicit activities in the world (TRAFFIC, 2008; Esmail 2 Museo Nacional de Ciencias Naturales-CSIC, José Gutiérrez et al., 2020), generating a turnover of around 20 billion Abascal 2, 28006 Madrid, Spain. dollars annually (Nellemann et al., 2016). 3 Research Unit of Biodiversity, Gonzalo Gutiérrez Quirós s/n, Both legal and illegal exotic pet trades are widely Oviedo University - Campus Mieres, Edificio de Investigación, considered as important mechanisms of intercontinental 33600 Mieres, Spain. and international pathogen pollution (e.g., Fisher 4 Catalonian Reptiles and Amphibians Rescue Centre, 08783 Barcelona, Spain. and Garner, 2007; O’Hanlon et al., 2018), including * Corresponding author. E-mail address: [email protected] the chytrids Batrachochytrium dendrobatidis (Bd; O’Hanlon et al., 2018), B. salamandrivorans (Bsal; © 2021 by Herpetology Notes. Open Access by CC BY-NC-ND 4.0. Martel et al., 2014) and ranaviruses (RV; Schloegel et 778 Barbora Thumsová et al. al., 2009). Both chytrid fungi and ranaviruses are listed thoroughly inspected by the police and the CITES by the World Organisation for Animal Health (Schloegel national department. Illegally acquired individuals et al., 2010; OIE, 2019) and therefore, in theory, control were confiscated and transported to the Catalonian measures by all countries importing amphibians must Reptiles and Amphibians Rescue Centre (CRARC). All be implemented to ensure that animals are free of these individuals were housed separately in tanks, and were pathogens (OIE, 2019). immediately checked visually for disease symptoms, The European Union (EU) has been documented as and sampled as described above. For the specimens a top importer of legally and illegally collected wild found dead, a toe-clip and a small piece of liver were animals (Engler and Parry-Jones, 2007). Within the also taken and stored in 70% ethanol. EU, Spain houses a sizable community of keepers, with DNA was extracted from swab samples using PrepMan extensive amphibian collections containing specimens Ultra reagent, and was extracted from tissue samples imported from all around the world (e.g., Fitzpatrick with the DNeasy Blood and Tissue Kit (Qiagen, Hilden, et al., 2018; Lastra-González et al., 2020). The largest Germany) following the manufacturer’s instructions. amphibian and reptile trade fairs of Southern Europe Real-time Taqman PCR assays were conducted take place in some of Spain’s main cities. Local traders, following the protocols suggested by Boyle et al. (2004) as well as traders from all over Europe, gather four for Bd, Blooi et al. (2013) for Bsal, and Leung et al. times a year at these events to sell a great variety of (2017) for RV. Samples were amplified on a MyGo exotic species. Despite the fact that imported exotic mini PCR machine in duplicate and against negative individuals may serve as vectors of both fungal and and positive controls with known concentrations of viral infections, sanitary regulations and legislation genomic equivalents of zoospores/virions. A sample was governing the practice to prevent pathogen spread in assigned as positive when the infection load was equal Spain are still not officially established. to or higher than 0.1 zoospores for Bd/Bsal or 3 virions Here we report the results of Bd, Bsal and RV testing for RV, and when the amplification curve presented a in four trade fairs that took place in Spain between 2017 robust sigmoidal shape. Where just one replicate of a and 2019, and also within a large illegal private collection sample was amplified, the sample was analysed a third recently confiscated in the centre of Barcelona city. time and considered positive only if the curve of the third amplification yielded a positive result. Materials and Methods Results Sampling at Spanish trade fairs took place between 2017–2019, three times in Madrid, and once in Over the four sampling events in trade fairs, a total of Barcelona. All pet traders and all amphibian species 82 individuals comprising 18 families and 34 species were sampled as described below. At least one sample from 20 traders were sampled (Table 1). Seventy-seven was collected per container when multiple animals were samples were tested for Bd, fifty-five for Bsal and housed together. Additionally, five imported terrapins forty-six for RV presence. While around 10% of the (two individuals of Graptemys pseudogeographica specimens tested positive for Bd and 2.2% of specimens (Gray, 1831), one individual of Cuora amboinensis tested positive for RV, no individuals tested positive for (Daudin, 1802), and two individuals of Kinosternon Bsal. Infected individuals were detected at all sampling baurii (Garman, 1891)) were sampled and tested for events. Bd infection was detected in individuals RV presence. The whole body of each individual was belonging to the families of (1/6 swabbed with a sterile cotton-tipped dry swab (MWE individuals), Arthroleptidae (1/2), Ceratophryidae medical wire, UK), and a new pair of disposable nitrile (2/9), Hylidae (1/1), Pipidae (2/3), and Pyxicephalidae gloves was used for each animal. Samples were stored (1/4). No Bd infected individuals presented disease dry and refrigerated until laboratory analyses within 1-2 symptoms. Ranavirosis symptoms were observed in weeks. three specimens of Litoria caerulea (White, 1790) In the spring of 2020, a police intervention was carried (Pelodryadidae), of which one individual also tested out in a private collection of urodele amphibians in positive for RV (Fig. 1A). All sampled terrapins were the centre of Barcelona city. The collection comprised negative for RV. Bd or RV infection was detected in a great number of exotic and Iberian-native urodele a total of five traders. One trader sold Bd-infected species. For the administrative and court proceedings, animals repeatedly across different trade fairs (3 out 4), the origin and legal status of all individuals were all of which were imported. Despite traders being very Incidence of emerging pathogens in the legal and illegal amphibian trade in Spain 779

Table 1. For each pathogen the number of positive animals and sample size are shown. The proportion of infected animals appears in brackets, and the positive species appear in the last column.

Family Bd Pos/N RV Pos/N Bsal Pos/N Positive species Ambystomatidae 1/6 (16.7) 0/2 0/5 Ambystoma mexicanum Arthroleptidae 1/2 (50.0) 0/0 0/2 Arthroleptis poecilonotus Bufonidae 0/4 0/2 0/3 Centrolenidae 0/1 0/0 0/1 Ceratophryidae 2/9 (22.2) 0/5 0/6 Ceratophrys cranwelli Lepidobatrachus laevis Dendrobatidae 0/28 0/19 0/19 Emydidae 0/0 0/2 0/0 Geoemydidae 0/0 0/1 0/0 Hylidae 1/1 (100.0) 0/1 0/1 Osteopilus spp. Hyperoliidae 0/4 0/3 0/3 Kinosternidae 0/0 0/2 0/0 Mantellidae 0/1 0/0 0/0 Pelodryadidae 0/4 1/4 (25.0) 0/4 Litoria caerulea Phyllomedusidae 0/4 0/0 0/2 Pipidae 2/3 (66.7) 0/3 0/3 Xenopus fraseri Pyxicephalidae 1/4 (25.0) 0/2 0/4 Pyxicephalus adspersus Rhacophoridae 0/2 0/0 0/1 0/4 0/0 0/1 TOTAL 8/77 1/46 0/55

accessible for sampling during trade fairs, none of them Laurenti, 1768, S. corsica Savi, 1838, Triturus carnifex were interested in further pathogen screening, even (Laurenti, 1768) and T. dobrogicus (Kiritzescu, 1903). those selling infected individuals, and also refused our Wild populations of 9 of these wild-caught (or suspected advice on how to treat the animals. to be) species are showing decreasing population

Twenty-nine species of urodele were found in the trends, and two of them are categorised as threatened private Barcelona urodele collection during the police by the IUCN (IUCN, 2020). Most of the specimens intervention. While most of the species (58.6%) had showed obvious signs of malnutrition and weakness. been acquired from the legal or illegal amphibian Four individuals were found dead, some of them in trade, up to 24.1% of the species had been harvested an advanced stage of decay (Fig. 1B). All individuals illegally directly from the wild. These included Spanish illegally acquired were confiscated, and the rescue autochthonous species protected by national regulations, centre received a total of 57 individuals of 11 species, such as arnoldi Carranza & Amat, 2005, C. dead specimens included. Four confiscated individuals asper (Dugès, 1852), Ichthyosaura alpestris (Laurenti, died within a couple of hours. Also, two legally acquired 1768), Lissotriton boscai (Lataste, 1879), L. helveticus specimens of Chinese Giant Andrias (Razoumowsky, 1789), Salamandra salamandra davidianus (Blanchard, 1871), protected by CITES (Linnaeus, 1758) and S. algira Bedriaga, 1883 (S. algira and listed on Appendix I (Table 2), were confiscated also being regulated by CITES; Table 2). The remaining due to their poor welfare, and ultimately deposited in 17.2% species were of unknown origin but probably Barcelona Zoo. Four confiscated individuals of three wild-caught, and also included four species listed on species (C. arnoldi, S. algira, S. salamandra) died Appendix II or III of the Bern Convention (S. atra within 24 hours. All sampled specimens (56) tested

 780 Barbora Thumsová et al.

Figure 1. (A) Ranavirosis symptoms observed in a legally traded specimen of Litoria caerulea during an amphibian and reptile fair trade. (B) Dead individual in an advanced stage of decay found in the same tank as a living individual of Aneides lugubris during the police intervention of a private urodele collection.

negative for RV and Bsal, but four specimens of I. Discussion alpestris and one specimen of L. boscai tested positive Our results indicate that amphibian pet trade may for Bd. One carcass of I. alpestris tested positive for Bd imperil amphibian conservation, both directly by (Table 2), while the other ones, which were in a very bad exploitation of wild populations, and indirectly by condition, tested negative. contributing to pathogen spread. A private collection confiscated by the police contained several illegally

Table 2. Urodele species of a private collection tested for the presence of Bd, Bsal and RV. Origin, probable origin of individuals: T, wildlife trade; WC, wild-caught; Legal status: Leg, kept legally; Illeg, kept illegally; National law, level of protection of Spanish native species according to the national regulation ‘Real Decreto 139/2011’: DE, danger of extinction; L, listed; VU, vulnerable; CITES, species category according to CITES appendices I and II (UNEP 2021); IUCN, conservation status according to IUCN Red List (IUCN 2020) and the Species+ website for CITES species (UNEP 2021): CR, critically endangered; NT, near threatened; LC, least concern; VU, vulnerable; N dead, number of individuals found dead or which died within 24 hours of confiscation. Bd Pos/N, Bd positive animals, sample size and proportion of infected animals. All specimens tested negative for Bsal and RV.

Species Origin Legal status National law CITES IUCN N dead Bd Pos/N Ambystoma mexicanum T Leg II CR 1 0/1 Andrias davidianus T Leg I CR 0 0/1 Calotriton arnoldi WC Illeg DE CR 1 0/10 Calotriton asper WC Illeg L NT 1 0/12 Ichthyosaura alpestris WC Illeg VU LC 1 4/6 (66.7) Lissotriton boscai WC Illeg L LC 0 1/2 (50.0) Lissotriton helveticus WC Illeg L LC 0 0/2 Notophthalmus viridescens T Leg LC 1 0/1 Plethodon hubrichti T Leg VU 0 0/1 Salamandra algira WC Illeg VU III VU 2 0/18 Salamandra salamandra WC Illeg LC 1 0/2

 Incidence of emerging pathogens in the legal and illegal amphibian trade in Spain 781 wild-caught species, many individuals presenting signs keeping can significantly impact animal welfare (Baker of poor welfare, and almost nine percent of individuals et al., 2013). was infected by Bd. More worryingly, we detected Bd The prevalence of Bd infection detected in our case and RV within legal amphibian trade fairs in Spain, study of an illegal collection (8.9%; Table 2) was, and traders selling infected animals showed no interest however, much lower than that detected in a similar study in further collaborations even when offered free of in Brazil (40.0%; De Paula et al., 2012). This difference charge. is most likely due to the relatively low susceptibility to The still growing amphibian trade poses a significant Bd of the adult stages of most urodeles represented in threat for many amphibian species and their conservation this collection (e.g., El Mouden et al., 2011; Obon et al., (Tapley et al., 2011). Intentional wild amphibian 2013; Martínez-Silvestre et al., 2020a). Alternatively, harvesting is one of the largest threats to species the lower incidence of Bd in our case study could be persistence and population survival (Stuart et al., 2004; due to the higher likelihood to obtain false negatives Schlaepfer et al., 2005; Chanson et al., 2008; Anderone when DNA extraction is performed by PrepMan Ultra et al., 2012). An increasing demand by collectors for reagent (Bletz et al., 2015). Fortunately, even though rare species is especially impacting endemic, protected the collection comprised of several Asiatic species, species, whose distribution is often restricted to small potential reservoirs of Bsal infection (UNEP-WCMC, ranges (Milner-Gulland, 2001; Webb et al., 2002; 2016), all confiscated individuals tested negative for Schlaepfer, et al., 2005). In our case study, some Bsal. However, chytridiomycosis outbreaks associated protected and highly range-restricted species such as C. with Bsal infection in Spanish captive amphibians asper and C. arnoldi were found, and it is likely that have been reported (Fitzpatrick et al., 2018), and the most of them were harvested illegally directly from the illegal collection in our study was located very close to wild. In fact, some confiscated individuals of C. arnoldi the only known Bsal outbreak outside the core area of were still marked with the visible implant elastomers North-Western Europe (Martel et al., 2020). Moreover, used in the conservation program of this species. Both some extremely thin and weak confiscated specimens of species are protected by the Spanish national regulations C. asper and C. arnoldi tested positive for Chlamydia (Real Decreto 139/2011; Decreto legislativo 2/2008), (Martínez-Silvestre et al., 2020b), a bacterium associated but are not regulated by CITES or the EU Wildlife with poor amphibian captive breeding (Densmore and Trade Regulation 338/97, the unique international Green, 2007; Martel et al., 2012). mechanisms regulating over-exploitation of wild fauna The overall proportion of infected individuals detected for the international trade (Janssen and Leupen, 2019; in trade fairs in our study (11.0%, Table 2) was only Altherr and Lameter, 2020). Unfortunately, once a slightly higher than that found in a trade fair in the species protected only in its country of origin is illegally Netherlands (9.0%, Spitzen-van der Sluijs et al., 2011), exported, any further trade and movement occurs but much lower than that detected in a frog market in without any legal persecution (Altherr and Lameter, South America comprising only wild-caught specimens 2020; Toland et al., 2020), and can be at serious risk of (100%, Catenazzi et al., 2010). However, in our case the being over-harvested (Toland et al., 2020). In this study, infection was confirmed in 25% of expositors, twice as this is particularly true for the critically endangered C. much as reported by Spitzen-van der Sluijs et al. (2011). arnoldi, restricted to only two fragmented population Whereas Bd infected individuals did not present any groups in the Montseny Natural Park in Catalonia. signs of the disease, evident symptoms of ranavirosis Even when amphibians’ ability to experience pain or were exhibited by some specimens, and yet they were develop complex behaviours is known (e.g., Burghardt, still offered for sale. 2013), only a few studies relating the impact of the The uncontrolled pet trade, even when it is legal, may wildlife trade to animal welfare have been published pose a high risk not only to wildlife, but also to public (Baker et al., 2013). Many specimens from the private health (Baker et al., 2013; Pasmans et al., 2017). In collection in our study showed obvious signs of response to Bsal emergence, the European Commission malnutrition and weakness, and some of them died took action to temporarily regulate the movement of within 24 hours of transportation to the rescue centre. captive urodeles, affecting both animal importation Dead individuals were found in the same tanks as into the EU, and intra-EU movement. All importers living individuals, some of them in an advanced stage now have to certify that urodele individuals have been of decay, confirming that wildlife trade and animal examined and are free of Bsal and symptoms of the 782 Barbora Thumsová et al. disease (EU 2018/320). Nevertheless, our findings Journal of Clinical Microbiology 51: 4173–4177. emphasise the persistent risk caused by the introduction Boyle, D.G., Boyle, D.B., Olsen, V., Morgan, J.A.T., Hyatt, A.D. of new pathogens, or more virulent strains of pathogens (2004): Rapid quantitative detection of chytridiomycosis (Batrachochytrium dendrobatidis) in amphibian samples using already present, through translocations of all captive real-time Taqman PCR assay. Diseases of Aquatic Organisms and wild-caught amphibians, not only urodele species. 60: 141–148. Considering that, in practice, Spain does not have any Burghardt, G.M. (2013): Environmental enrichment and cognitive sanitary and control measures established for traded complexity in reptiles and amphibians: Concepts, review, and individuals with respect to Bd and RV, implementation implications for captive populations. Applied Animal Behaviour of such measures could be of utmost importance Science 147: 286–298. (Pasmans et al., 2017). Private keepers and traders Catenazzi, A., Vredenburg, V.T., Lehr, E. (2010): Batrachochytrium dendrobatidis in the live frog trade of Telmatobius (Anura: should be profoundly interested in maintaining strict Ceratophryidae) in the tropical Andes. Diseases of Aquatic biosafety practices to prevent, at the very least, disease Organisms 92: 187–191. outbreaks within their collections and stocks. Ceballos, G., Ehrlich, P.R., Barnosky, A.D., García, A., Pringle, R.M., Palmer, T.M. (2015): Accelerated modern human-induced Acknowledgements. We thank the Forest Agents of Comunidad species losses: Entering the sixth mass extinction. Science de Madrid, V. Cadenas and A. Tarragó (Generalitat de Catalunya) Advances 1: 1–5. for assistance during sampling events, C. Monsalve-Carcaño and Chanson, J., Hoffmann, M., Cox, N., Stuart, S. (2008): The State C. Sausor for sampling and laboratory assistance, and J. Soler, of the World’s Amphibians. In: Threatened amphibians of the Z. Alonso and T. Ugrinović for assistance on maintenance of world, p.33–52. Stuart, S., Hoffmann, M., Chanson, J., Cox, confiscated individuals. Comunidad de Madrid and Generalitat N., Berridge, R., Ramani, P., Young, B., Ed. Lynx, Barcelona; de Catalunya extended permits for sampling at trade fairs. We IUCN, Gland, Switzerland; and Conservation International, thank G. Stride for reviewing the English. Arlington, Virginia, USA. Collins, J.P., Crump, M.L. (2009): Extinction in our Times. Global References Amphibian Decline. Oxford University Press, Oxford. Cooney, R., Kasterine, A., MacMillan, D., Milledge, S., Nossal, Altherr, S., Lameter, K., (2020): The rush for the rare: reptiles and K., Roe, D., et al. (2015): The trade in wildlife: A framework amphibians in the European pet trade. Animals 10: 2085. to improve biodiversity and livelihood outcomes, International Andreone, F., Loarie, S.R., Pala, R., Luiselli, L.M., Carpenter, Trade Centre, Geneva, Switzerland. A.I. (2012): Trade and exploitation of amphibians and reptiles: Courchamp, F., Angulo, E., Rivalan, P., Hall, R.J., Signoret, L., a conservation overview. Atti dell’Accademia delle Scienze di Bull, L., et al. (2006): Rarity value and species extinction: The Torino-Atti Classe di Scienze Fisiche 146: 85–93. anthropogenic Allee effect. PLoS Biology 4: 2405–2410. Auliya, M., Altherr, S., Ariano-Sanchez, D., Baard, E.H., Brown, De Paula, C.D., Pacífico-Assis, E.C., Catão-Dias, J.L. (2012): C., Brown, R.M., et al. (2016b): Trade in live reptiles, its impact Batrachochytrium dendrobatidis in amphibians confiscated from on wild populations, and the role of the European market. illegal wildlife trade and used in an ex situ breeding program in Biological Conservation 204: 103–119. Brazil. Diseases of Aquatic Organisms 98: 171–175. Auliya, M., García-Moreno, J., Schmidt, B.R., Schmeller, D.S., Densmore, C.L., Earl-Green, D. (2007): Diseases of Amphibians. Hoogmoed, M.S., Fisher, M.C., et al. (2016a): The global ILAR J. 48: 235–245. amphibian trade flows through Europe: the need for enforcing El Mouden, E.H., Slimani, T., Donaire, D., Fernández-Beaskoetxea, and improving legislation. Biodiversity and Conservation 25: S., Fisher, M.C., Bosch, J. (2011): First record of the chytrid 2581–2595. fungus Batrachochytrium dendrobatidis in North Africa. Baker, S.E., Cain, R., Kesteren, F. Van, Zommers, Z.A., D’Cruze, Herpetological Review 42: 71–75. N., MacDonald, D.W. (2013): Rough trade: Animal welfare in Engler, M., Parry-Jones, R. (2007): Opportunity or Threat: The the global wildlife trade. BioScience 63: 928–938. Role of the European Union in Gobal Wildlife Trade. TRAFFIC Blaustein, A.R., Han, B.A., Relyea, R.A., Johnson, P.T.J., Buck, Europe, Brussels, Belgium. J.C., Gervasi, S.S., et al. (2011): The complexity of amphibian Esmail, N., Wintle, B.C., t Sas-Rolfes, M., Athanas, A., Beale, population declines: Understanding the role of cofactors in C.M., Bending, Z., et al. (2020): Emerging illegal wildlife trade driving amphibian losses. Annals of the New York Academy of issues: A global horizon scan. Conservation Letters 13: e12715. Sciences 1223: 108–119. Fisher, M.C., Garner, T.W.J. (2007): The relationship between the Bletz, M.C., Rebollar, E.A., Harris, R.N. (2015): Differential emergence of Batrachochytrium dendrobatidis, the international efficiency among DNA extraction methods influences detection trade in amphibians and introduced amphibian species. Fungal of the amphibian pathogen Batrachochytrium dendrobatidis. Biology Reviews 21: 2–9. Diseases of Aquatic Organisms 113: 1–8. Fitzpatrick, L.D., Pasmans, F., Martel, A., Cunningham, A.A. Blooi, M., Pasmans, F., Longcore, J.E., Spitzen-Van Der Sluijs, A., (2018): Epidemiological tracing of Batrachochytrium Vercammen, F., Martel, A. (2013): Duplex real-Time PCR for salamandrivorans identifies widespread infection and associated rapid simultaneous detection of Batrachochytrium dendrobatidis mortalities in private amphibian collections. Scientific Reports and Batrachochytrium salamandrivorans in amphibian samples. 8: 13845. Incidence of emerging pathogens in the legal and illegal amphibian trade in Spain 783

IUCN (2020): The IUCN Red List of Threatened Species. Version OIE (World Organisation for Animal Health) (2019): Aquatic 2020-3. https://www.iucnredlist.org. Accessed on 15 of February Animal Health Code. http://www.oie.int/en/international- 2021. standard-setting/aquatic-code/access-online/. Accessed on 10 of Janssen, J., Leupen, B.T.C. (2019): Traded under the radar: Poor February 2021. documentation of trade in nationally-protected non-CITES Pasmans, F., Hellebuyck, T., Martel, A., Bogaerts, S., Braeckman, species can cause fraudulent trade to go undetected. Biodiversity J., Cunningham, A.A., et al. (2017): Future of keeping pet and Conservation 28: 2797–2804. reptiles and amphibians: Towards integrating animal welfare, Lastra-González, D., Baláž, V., Chajma, P., Vojar, J. (2020): human health and environmental sustainability. Veterinary Surveying for Batrachochytrium salamandrivorans presence in Record 181: 450. Spanish captive collections of amphibians. Diseases of Aquatic Rowley, J., Brown, R., Bain, R., Kusrini, M., Inger, R., Stuart, B., Organisms 142: 99–103. et al. (2010): Impending conservation crisis for Southeast Asian Leung, W.T.M., Thomas-Walters, L., Garner, T.W.J., Balloux, amphibians. Biology Letters 6: 336–338. F., Durrant, C., Price, S.J. (2017): A quantitative-PCR based Scheffers, B.R., Oliveira, B.F., Lamb, I., Edwards, D.P. (2019): method to estimate ranavirus viral load following normalisation Global wildlife trade across the tree of life. Science 366: 71– by reference to an ultraconserved vertebrate target. Journal of 76. Virological Methods 249: 147–155. Schlaepfer, M.A., Hoover, C., Dodd, K. (2005): Challenges in Martel, A., Adriaensen, C., Bogaerts, S., Ducatelle, R., Favoreel, evaluating the impact of trade in amphibians and reptiles on wild H., Crameri, S., et al. (2012): Novel chlamydiaceae disease in populations. BioScience 55: 256–264. captive . Emerging Infectious Diseases 18: 1020– Schloegel, L.M., Daszak, P., Cunningham, A.A., Speare, R., Hill, B. 1022. (2010): Two amphibian diseases, chytridiomycosis and ranaviral Martel, A., Blooi, M., Adriaensen, C., Rooij, P. Van, Beukema, disease, are now globally notifiable to the World Organization W., Fisher, M.C., et al. (2014): Recent introduction of a chytrid for Animal Health (OIE): An assessment. Diseases of Aquatic fungus endangers Western Palearctic salamanders. Science 346: Organisms 92: 101–108. 630–631. Schloegel, L.M., Picco, A.M., Kilpatrick, A.M., Davies, A.J., Martel, A., Vila-Escale, M., Fernández-Giberteau, D., Martinez- Hyatt, A.D., Daszak, P. (2009): Magnitude of the US trade in Silvestre, A., Canessa, S., van Praet, S., et al. (2020): Integral amphibians and presence of Batrachochytrium dendrobatidis chain management of wildlife diseases. Conservation Letters and ranavirus infection in imported North American bullfrogs 13: 1–6. (Rana catesbeiana). Biological Conservation 142: 1420–1426. Martínez-Silvestre, A., Obón, E., Tarrago, A. (2020b): Primer Skerratt, L.F., Berger, L., Speare, R., Cashins, S., McDonald, diagnóstico de Chlamydia en tritón del Pirineo (Calotriton K.R., Phillott, A.D., et al. (2007): Spread of chytridiomycosis asper) y tritón del Montseny (Calotriton arnoldi) vinculada has caused the rapid global decline and extinction of frogs. al coleccionismo ilegal de anfibios. Boletín de la Asociación Ecohealth 4: 125–134. Herpetológica Española 31: 173–176. Spitzen-van der Sluijs, A.M., Martel, A., Wombwell, E., Rooij, P. Martínez-Silvestre, A., Trochet, A., Calvez, O., Poignet, M., Le Van, Zollinger, R., Woeltjes, T., et al. (2011): Clinically healthy Chavalier, H., Souchet, J., et al. (2020a): Presence of the fungus amphibians in captive collections and at pet fairs: A reservoir Batrachochytrium dendrobatidis, but not Batrachochytrium of Batrachochytrium dendrobatidis. Amphibia-Reptilia 32: salamandrivorans, in wild Pyrenean Brook (Calotriton 419–423. asper) in Spain and France. Herpetological Review 51: 738– Stuart, S.N., Chanson, J.S., Cox, N.A., Young, B.E., Rodrigues, 743. A.S.L., Fischman, D.L., Waller, R.W. (2004): Status and trends Milner-Gulland, E.J. (2001): The exploitation of spatially structured of amphibian declines and extinctions worldwide. Science 306: populations. In: Conservation of Exploited Species, p.87–109. 1783–1786. Reynolds, J.D., Mace, G.M., Redford, K.H., Robinson, J.G., Ed., Tapley, B., Griffiths, R.A., Bride, I. (2011): Dynamics of the trade Cambridge (United Kingdom), Cambridge University Press. in reptiles and amphibians within the United Kingdom over a Natusch, D.J.D., Lyons, J.A. (2012): Exploited for pets: The ten-year period. Herpetological Journal 21: 27–34. harvest and trade of amphibians and reptiles from Indonesian Toland, E., Bando, M., Hamers, M., Cadenas, V., Laidlaw, R., New Guinea. Biodiversity and Conservation 21: 2899–2911. Martínez-Silvestre, A., van der Wielen, P. (2020): Turning Nellemann, C., Henriksen, R., Kreilhuber, A., Stewart, D., negatives into positives for pet trading and keeping: A review of Kotsovou, M., Raxter, P., et al. (2016): The rise of environmental positive lists. Animals 10: 1–45. crime – a growing threat to natural resources peace, development TRAFFIC (2008): What’s driving the wildlife trade? A review of and security. UNEP-INTERPOL. expert opinion on economic and social drivers of the wildlife Obon, E., Carbonell, F., Valbuena-Ureña, E., Alonso, M., Larios, trade and trade control efforts in Cambodia, Indonesia, Lao R., Fernández-Beaskoetxea, S., et al. (2013): Chytridiomycosis PDR and Vietnam. East Asia and Pacific Region Sustainable surveillance in the critically endangered Montseny brook , Development Discussion Papers. East Asia and Pacific Calotriton arnoldi, northeastern Spain. Herpetological Journal Region Sustainable Development Department, World Bank, 23: 237–240. Washington, DC. O’Hanlon, S.J., Rieux, A., Farrer, R.A., Rosa, G.M., Waldman, B., UNEP (2021): The Species+ Website. Nairobi, Kenya. Compiled Bataille, A., et al. (2018): Recent Asian origin of chytrid fungi by UNEP-WCMC, Cambridge, UK. https://www.speciesplus. causing global amphibian declines. Science 360: 621–627. net. Accessed on 15 of February 2021. 784 Barbora Thumsová et al.

UNEP-WCMC (2016): Review of the risk posed by importing Asiatic species of Caudata amphibians (salamanders and newts) into the EU. UNEP-WCMC, Cambridge. Webb, J.K., Brook, B.W., Shine, R. (2002): Collectors endanger Australia’s most threatened snake, the broad-headed snake Hoplocephalus bungaroides. Oryx 36: 170–181. Wilson, K., Fenton, A., Tompkins, D. (2019): Wildlife Disease Ecology: Linking Theory to Data and Application. Cambridge, Cambridge University Press.

Accepted by Wouter Beukema